Další formáty:
BibTeX
LaTeX
RIS
@article{492938, author = {Vytisková, Soňa and Van Dyck, Sigrid and Van Schepdael, Ann and Hoogmartens, Jos and Glatz, Zdeněk}, article_location = {Netherlands}, article_number = {(1+2)}, keywords = {Capillary electrophoresis; enzymes; EMMA; sulfotranferase}, language = {eng}, issn = {0021-9606}, journal = {Journal of Chromatography A}, title = {Study of enzyme kinetics of phenol sulfotransferase by electrophoretically mediated microanalysis}, volume = {1032}, year = {2004} }
TY - JOUR ID - 492938 AU - Vytisková, Soňa - Van Dyck, Sigrid - Van Schepdael, Ann - Hoogmartens, Jos - Glatz, Zdeněk PY - 2004 TI - Study of enzyme kinetics of phenol sulfotransferase by electrophoretically mediated microanalysis JF - Journal of Chromatography A VL - 1032 IS - (1+2) SP - 319-326 EP - 319-326 PB - Elsevier SN - 00219606 KW - Capillary electrophoresis KW - enzymes KW - EMMA KW - sulfotranferase N2 - Electrophoretically mediated microanalysis (EMMA) was applied for the study of the kinetic parameters of the enzymatic reaction of phenol sulfotransferase SULT1A1 isoenzyme with 4-nitrophenol as a substrate. The SULT1A1 activity was determined by the quantitation of the product, 4-nitrophenyl sulfate, at 274 nm by using different injection and separation steps. This new approach solved the problem of the presence of the very strong inhibitor, adenosine 3',5'-bisphosphate (PAP), in the co-substrate solution (adenosine 3'-phosphate 5'-phosphosulfate, PAPS) which is unstable at room temperature. The inhibitor PAP was electrophoretically separated from the co-substrate PAPS before the injection of enzyme and substrate inside the capillary (and thus before their in-capillary encountering). With the developed in-capillary SULT1A1 activity assay an average Michaelis constant (Km) for 4-nitrophenol was calculated to be 0.84 M, a value which is consistent with a previously reported value. Strong substrate inhibition (above a 4-nitrophenol concentration of 2.5 M) was observed, and this is also in accordance with literature values. ER -
VYTISKOVÁ, Soňa, Sigrid VAN DYCK, Ann VAN SCHEPDAEL, Jos HOOGMARTENS a Zdeněk GLATZ. Study of enzyme kinetics of phenol sulfotransferase by electrophoretically mediated microanalysis. \textit{Journal of Chromatography A}. Netherlands: Elsevier, 2004, roč.~1032, (1+2), s.~319-326. ISSN~0021-9606.
|